In the use of genetic manipulation techniques in microorganisms, the genus Bacillus has, after E. coli, in recent years also formed the subject of extensive investigation. See, for example, Dubnau, in Experimental Manipulation of Gene Expression, Academic Press, (1983) 33-51 and Doi, Biotechnology and Genetic Engineering (1984) 2:126-155. Bacilli have now been used for a long time in the fermentation industry. Bacilli offer numerous advantages, such as good growth on inexpensive base materials, and in contrast to E. coli, do not produce any endotoxins. Furthermore, Bacilli are capable of secreting proteins into the growth medium, in particular, certain types of enzymes such as proteases and amylases, frequently produced in large amounts by Bacilli. These enzymes may be relatively inexpensively and conveniently isolated from the fermentation medium.
Because of the attractiveness of Bacilli as a host for the production of homologous or heterologous peptides, it is of substantial commercial interest to be able to make use of particular sequences associated with transcriptional and translational regulation, which would allow for efficient expression and secretion of the peptides of interest. There is, therefore, substantial interest in ways for isolating and analyzing these sequences from Bacillus or other sources, which would allow for the efficient screening of the sequences.